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Todd Boulevard, Nashville, TN 37208, USA; 7Systems Biology and Bioinformatics Group, University of Rostock, Universitätsplatz 1, 18055 Rostock, Germany; 8School of Computer Science, Carl

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Gilles Clermont 1 , Charles Auffray 2 , Yves Moreau 3 , David M Rocke 4 ,

Address: 1Department of Critical Care Medicine and CRISMA laboratory, University of Pittsburgh School of Medicine, Scaife 602,

3550 Terrace, Pittsburgh, PA 15261, USA; 2Functional Genomics and Systems Biology for Health, CNRS Institute of Biological Sciences,

7, rue Guy Moquet, BP8 94801 Villejuif Cedex, France; 3K.U Leuven, ESAT/SCD, Kasteelpark Arenberg 10, B-3001 Leuven-Heverlee, Belgium; 4Department of Public Health Sciences, University of California, Davis, One Shields Ave, Davis, CA 95616, USA; 5Department of Computer Science and Engineering Chalmers and Göteborg University, SE 41296, Göteborg, Sweden; 6Department of Surgery, Meharry Medical College, 1005 Dr D.B Todd Boulevard, Nashville, TN 37208, USA; 7Systems Biology and Bioinformatics Group, University of Rostock, Universitätsplatz 1, 18055 Rostock, Germany; 8School of Computer Science, Carleton University, 1125 Colonel By Drive, Ottawa, Ontario K1S 5B6, Canada; 9Computational Biology Unit Molecular Biotechnology Center, University of Torino, Via Nizza 52, I, 10126 Torino, Italy; 10Institutionen för Medicin, Karolinska Universitetssjukhuset, Solna, 171 76 Stockholm, Sweden; 11Computational Medicine Center, University of Cincinnati, 3333 Burnet Avenue, Cincinnati, OH 45229, USA; 12Department of Electrical Engineering and Computer Science, College of Engineering, University of Tennessee, 1122 Volunteer Boulevard, Knoxville, TN 37996, USA; 13The Unit for Clinical Systems Biology, The Queen Silvia Children’s Hospital, Gothenburg 40530, Sweden

Corresponding author: Gilles Clermont, cler@pitt.edu

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Ab bssttrraacctt

Systems biology has matured considerably as a discipline over the last decade, yet some of the

key challenges separating current research efforts in systems biology and clinically useful results

are only now becoming apparent As these gaps are better defined, the new discipline of systems

medicine is emerging as a translational extension of systems biology How is systems medicine

defined? What are relevant ontologies for systems medicine? What are the key theoretic and

methodologic challenges facing computational disease modeling? How are inaccurate and

incomplete data, and uncertain biologic knowledge best synthesized in useful computational

models? Does network analysis provide clinically useful insight? We discuss the outstanding

difficulties in translating a rapidly growing body of data into knowledge usable at the bedside

Although core-specific challenges are best met by specialized groups, it appears fundamental that

such efforts should be guided by a roadmap for systems medicine drafted by a coalition of

scientists from the clinical, experimental, computational, and theoretic domains

Published: 29 September 2009

Genome Medicine 2009, 11::88 (doi:10.1186/gm88)

The electronic version of this article is the complete one and can be

found online at http://genomemedicine.com/content/1/9/88

Received: 15 May 2009 Revised: 11 June 2009 Accepted: 15 September 2009

© 2009 Clermont et al.; licensee BioMed Central Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited

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Co orrrre essp ponden ncce e

Recent years have seen the rise of systems biology as a

legitimate discipline Although consensus exists about what

the fundamental tools are (high-throughput data from

several biologic scales, high-definition imaging, and

compu-tational modeling), no such consensus exists as to what

defines the broad agenda of systems biology A growing

awareness is found that, despite such major technologic advances, fundamental obstacles separate systems biology from clinical applications Bridging these gaps will require a focused and concerted effort What defines systems medicine

as a discipline? What should it seek to accomplish? How should knowledge from disparate sources be assembled into ontologies relevant to systems medicine? How are multiscale

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data to be synthesized by corresponding multiscale models?

What is the burden of proof that such models are valid and

predictive of clinically relevant outcomes? Is network

analysis a useful tool for systems medicine?

Physicians, basic scientists, mathematicians, statisticians

and computer scientists met at the Third Bertinoro Systems

Biology workshop [1], sponsored by the University of

Bologna, focused on the theme ‘Systems Biology Meets the

Clinic’ to address these questions Participants sought to

identify key challenges facing the successful translation of

systems biology to the clinical arena and discussed and

debated a roadmap seeking to address them The meeting,

held over a 4-day period, comprised plenary lectures followed

by extensive thematic discussions, formal and informal,

centered on the theme of systems medicine as a distinct

translational discipline [2]

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De effiin niin ngg ssyysstte em mss m me ed diicciin ne e

Workshop participants proposed that systems medicine be

defined as the application of systems biology to the

prevention of, understanding and modulation of, and

recovery from developmental disorders and pathologic

processes in human health Although no clear boundary

exists between systems biology and systems medicine, it

could be stated that systems biology is aimed at a

funda-mental understanding of biologic processes and ultimately at

an exhaustive modeling of biologic networks, whereas

systems medicine emphasizes that the essential purpose and

relevance of models is translational, aimed at diagnostic,

predictive, and therapeutic applications Accordingly,

advances in systems medicine must be assessed on both a

medical and more basic biologic scale, as the

correspon-dence between medicine and biology is intricate Some

seemingly straightforward biologic models may have an

important medical impact, although some impressively

complex molecular models may not be immediately

medically relevant Whereas systems biology may have so far

focused primarily on the molecular scale, systems medicine

must directly incorporate mesoscale clinical information

into its models; in particular, classic clinical variables,

biomarkers, and medical imaging data As an example, it has

become increasingly clear that prognostic and predictive

models for malignant tumors using expression data cannot

ignore information from classic prognostic indices [3]

Furthermore, because of the necessary multiscale nature of

the models bridging embedded levels of organization from

molecules, organelles, cells, tissues, organs, and all the way

to individuals, environmental factors, populations, and

ecosystems, systems medicine aims to discover and select

the key factors at each level and integrate them into models

of translational relevance, which include measurable

readouts and clinical predictions Such an approach is

expected to be most valuable when the execution of all

experi-ments necessary to validate sufficiently detailed models is limited by time, expenses (e.g., in animal models), or basic ethical considerations (e.g., human experimentation) Systems medicine as a discipline did not emerge from clinical medicine, but draws its relevance from it Conversely, advances in systems biology created the necessary conditions and tools for the emergence of systems medicine

Accordingly, although it may be appropriate to position systems medicine as an extension of systems biology from a historical perspective, the former also draws from several other disciplines, such as clinical medicine and population epidemiology, less familiar to systems biologists

S Sccaalle e ssp pe ecciiffiicc m mo od de elliin ngg vve errssu uss m mu ullttiissccaalle e m mo od de elliin ngg Computational models have for the most part attempted to assimilate massive data streams collected by using global measurement technologies (techniques that look at the complete set of genes, transcripts, proteins, metabolites, or other features in an organism) by using high-throughput techniques and have been, by and large, scale specific Such attempts target the development of predictive mathematic and computational models of functional and regulatory biologic networks Specific biologic hypotheses can thus be tested by designing a series of relevant perturbation experiments [4] Clear merit inheres in such an incremental approach, yet its true potential is likely to be realized only when such data-driven, bottom-up approaches are com-bined with top-down, model-driven approaches to generate new medically relevant knowledge

An open question is whether integrative systems-biology approaches can reveal underlying principles related to the aforementioned biologic functions It is probably improper

to speak of the existence of biologic laws in the sense of physical laws, yet probably deeper dynamic principles guide the evolution of biologic systems Energetic and physical constraints play an important role in all scale-specific models Additional principles at play across multiple scales

in biologic systems are far less apparent Thus, it appears prudent at this stage that top-down and multiscale models seek to recapitulate scale-specific observables As mentioned previously, if computational models are to be validated by experiments such as randomized clinical trials and become predictive of therapeutic interventions, relevant system observables must be included

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On ntto ollo oggiie ess rre elle evvaan ntt tto o ssyysstte em mss m me ed diicciin ne e Considerable attention should be paid to the development of ontologies relevant to systems medicine Such ontologies must reflect knowledge based on biologic function, rather than on biologic structure Indeed, structure is permissive to function, and clearly, a wide variety of structures could have evolved, under genetic, molecular, or physical constraints to

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accomplish a given function Examples include energy

generation and storage and transmission of information

The recent emphasis on mapping structure into function is

vital to the advancement of systems medicine In addition, it

appears that the development of appropriate ontologies

could promote a (re)interpretation of empiric evidence in

light of such ontologies As an example, experimental data

often appear to support contradictory hypotheses of limited

scope, when in fact the evidence can be reconciled under a

broader synthesis of the evidence

Progress in developing meaningful ontologies for systems

medicine will challenge our current intuition of the nature of

a biologic function Recent efforts at data reduction for

longitudinal expression data, by using principal-component

analysis to identify and monitor health and disease

“trajec-tories”, represent an attempt at understanding such

“eigen-processes” from a data-driven perspective [5,6] Typically

and unfortunately, such processes have limited intuitive

meaning when interpreted through the prisms of currently

existing ontologies Alternatively, existing community (for

example, Gene Ontology (GO)) or commercial efforts aimed

at developing a phenotype-driven ontology (e.g., annotating

genes to a priori defined functions such as “cell-cycle” or

“inflammatory response”) are commendable and clearly of

great value, although it is apparent that extensive

cross-contamination exists between such functional assignments

and the response to even the simplest experimental

pertur-bation of functions Knowledge representations relevant to

systems medicine will probably lie within this spectrum, and

computational efforts will likely be crucial to their

development

Both data-driven techniques and simulation-based

tech-niques open possibilities of reinterpreting what is meant by

biologic function, yielding new knowledge representations

Multiscale models that include phenotypes as inputs or

readouts will provide mechanistic insight into the dynamic

interplay of such redefined functions, and plausibly suggest

phenotypically based therapeutic targets

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Ne ew w k kn no ow wlle ed dgge e aan nd d ffaallsse e d diisscco ovve erryy

Experimental design and statistical analysis should be dealt

with rigorously, as they play essential roles in discovery and

validation in systems biology and medicine [7] Study design

is often the weakest point of complex molecular studies in

systems biology and medicine For example, patients with a

disease such as ovarian cancer may be compared with

normal controls to discern aberrant regulation of pathways

If controls are not carefully selected to be comparable with

patients demographically and in other covariates (age, sex,

income, social class), then differences observed may be

attributable to factors other than the disease

Researchers are often unduly optimistic about sample sizes required to show differences, and they fail to consider many confounding effects Interindividual variability in humans can be large, often the largest effect in a study This provides

an avenue for exploration of individual effects, leading to personalized medicine, but also can make detection of differences across subjects quite difficult

High-throughput technologies have introduced new challenges to experimental design and interpretation of results Avoiding false positives may result in difficulties in identifying true positive Standard approaches to correcting for multiple-testing on datasets generated by global analysis, such as expression microarray, rely on the incorrect assumption that each value is independent of other values More recent approaches do not fully resolve this problem [8] Greatly increasing sample sizes is generally impractical

A more practical approach is to make increased use of a priori biologic knowledge, either by trimming the list of analytes to a relatively small number for which the multiple-testing correction is modest, or by multiple-testing pathways or groups of genes [9] This is usually done not by testing every group of genes defined by a GO term or a Kyoto Encyclo-pedia of Genes and Genomes (KEGG) pathway, but by selectively testing those thought to be of importance Because this more-focused approach, in its effort to improve specificity, is ontology dependent, it may bear a subjective element as to the certainty of prior knowledge It, therefore, also carries the risk of lacking sensitivity

Addressing the previously mentioned challenges may have direct clinical implications A frequent problem encountered

by clinicians is that patients appearing to have the same disease may not respond to the same treatment Some patients even experience severe adverse effects from the treatment Variable treatment response is also one of the most important causes of the huge costs involved in drug development Taken together, these cause both increased suffering and costs Ideally, physicians should be able, routinely and noninvasively, to measure a few diagnostic biomarkers to personalize medication for each patient At present, not enough knowledge exists about the causes for variable treatment responses in most common diseases However, recent studies of genetic markers for response to treatment with anticoagulants indicate that personalized dosage may become a clinical reality within the next 5 to

10 years [10] The main problems involved in finding markers for personalized dosage are that each complex disease may involve altered interactions between hundreds

or thousands of genes that can differ among patients This heterogeneity may, in turn, depend on both genetic and environmental factors In addition to this complexity, significant problems are involved in clinical research Ideally, a study aiming to find markers for personalized medication would involve a known external cause, a key cell

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type, and a read-out, all of which can be studied

experi-mentally in patient samples

For most complex diseases, all of these factors are not

readily available It is therefore important to find model

diseases, in which all those factors can be studied together in

patient samples by using high-throughput technologies and

systems biologic principles [11] Such model diseases might

be used to develop and apply the methods required to find

markers for personalized medicine

It also has been suggested that the same methods might be

applied to find markers to predict the risk of developing

disease [12] If successful, this may lead to a new era of

preventive medicine Finally, the methods may be of great

value for drug development If it were possible to predict

which patients respond to medication, this would result in

increased efficacy and reduced risk of not being able to market

drugs that have been developed at great cost Conversely,

delineation of patients that do not respond to a medication

may help to develop new drugs for that specific subgroup We

suggest that acute inflammatory diseases, such as severe

trauma, sepsis, and pancreatitis, might be very attractive test

beds for the development of such methods Similarly, chronic

ailments, such as diabetes and other autoimmune disorders,

meet several of the criteria mentioned earlier and are of

prominent clinical and societal relevance

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Ne ettw wo orrk k aan naallyyssiiss

A network represents a set of objects and their mutual

relations Much biologic and medical knowledge can be

naturally represented as networks: protein-interaction

networks, metabolic networks, gene co-expression networks,

disease networks, and many more Growing concerns regard

current trends in network analysis in systems biology and

potential extension to the clinical arena through the

construction of “diseasomes” [13] Do network

representa-tions actually convey new knowledge, or are they just a

convenient and eye-catching way to represent data? How

can such networks be used to extract new information that is

relevant to understanding biologic systems and guiding

clinical practice? Are current approaches adequately

repre-senting the types of entities and the specific nature of their

relations that determine disease pathophysiologic processes?

What challenges might be resolved and opportunities

opened for both basic research and clinical practice if

standards could be broadly adopted in our knowledge

representation, data collection, publication, and reasoning,

and if fundamental chemical, physical, and biologic entities

and processes could be included in network representations?

How might this be enabled by the adoption of

disease-oriented ontologies? From a mathematic and computational

perspective, what topologic, dynamic, and conditional

properties could allow the identification of the nodes in a

network whose perturbation would yield adversely affected

or clinically improved biologic states?

Although the methods used to analyze networks might still be primitive, they are already providing useful information, especially on the genetics of disease It is now possible to integrate information from various biologic networks to identify genes involved in both mendelian and complex diseases In such research efforts, careful thought must be given to how network inferences from microarray and other types of data are evaluated The development of such tools should ideally involve an open dialogue between experi-mentalists, modelers, and clinicians, who should be able to assess tools best suited to their application A need exists for systematic benchmark testing and comparative evaluation of the major tools available For example, current methods tend

to focus more on testing performance capabilities over simulated data or for functional enrichment in GO categories that may not be very relevant to clinically relevant phenomena The identification of both disease-causative genes and potential therapeutics has begun to be approached by using integrative network-relevant methods for knowledge representation and reasoning [14,15] Another possibility is the identification of specific interactions that have been extensively validated, a so-called ‘gold standard’ for the identification of causal, mechanistic, and deterministic factors in a complex network Some of these issues have been raised within the Dialogue on Reverse Engineering Assessment and Methods (DREAM) initiative [16] For example, representing gene interactions with graph algorithms may be a useful method to discover parts of a network that are not fully resolved [17] The biologic plausibility of such representations could then be integrated with other technologies and discussed with basic biologists and clinicians Another approach is to extend network analysis to evaluate disease-specific ontologies [18]

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Co on nccllu ussiio on nss aan nd d rre ecco om mm me endaattiio on nss

We consider that improvements in academic infrastructure are sorely needed to facilitate cross-disciplinary trans-lational studies that can someday connect what can be learned by using model organisms with real-time samples from patients Such improvements include, but are not limited to sufficient funding, appropriate development of mechanisms allowing academic recognition of all partici-pants of transdisciplinary teams, the creation of centers of excellence in systems medicine and specific training programs, and enhancement of the attractiveness of a medical career for individuals with training in quantitative fields Recognition of systems medicine in the clinical arena should be promoted at the professional society and journal editorial levels Indeed, whereas bioinformatics exercises can access mainstream clinical literature on account of the value of a significance test, the burden of proof appears

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disproportionately higher for computational disease and

therapeutic models of clinical relevance Additionally, the

construction of a roadmap for systems medicine, facilitated

by enhanced visibility in the more clinically oriented medical

literature, will be essential to chart effort and progress We

present essential elements of such a roadmap, as well as

underlying rationale (Figure 1)

A serious and useful dialogue between the clinic and systems

biology has begun We hope that future developments will

provide continuing evidence that the systems-biology

community has taken this development to its heart, building

systems medicine on a millennium of scholarship and

medical tradition

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Ab bb brre evviiaattiio on nss

DREAM = dialogue on reverse engineering assessment and

methods; GO = gene ontolology; KEGG = Kyoto

Encyclo-pedia of Genes and Genomes

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Co om mp pe ettiin ngg iin ntte erre essttss The authors declare that they have no competing interests

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Au utth ho orrss’’ cco on nttrriib bu uttiio on nss All authors contributed text on their specific domains of expertise GC collated text All authors reviewed the assembled text for accuracy

A Acck kn no ow wlle ed dgge emen nttss

We thank Michael Langston, Devdatt Dubhashi, and Mikael Benson for organizing the Bertinoro Systems Biology workshop, and the Bertinoro University Center, University of Bologna, for their generous support of the workshop

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Fiigguurree 11

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